Differential turnover of myosin chaperone UNC-45A isoforms increases in metastatic human breast cancer.

UNC-45A is a molecular chaperone targeted to non-muscle myosins and is essential for cell division. Here, we show that UNC-45A mRNA and protein expression was elevated in human breast carcinomas and cell lines derived from breast carcinoma metastases. Moreover, small hairpin RNA knockdowns of endogenously overexpressed UNC-45A in the most metastatic cell line led to significant decreases in the rates of cell proliferation and invasion, concomitant with reduction in the interaction of myosin II with actin filaments. Exploring the mechanism of these findings further, we found that UNC-45A is alternatively expressed at the mRNA and protein levels as two isoforms. The two isoforms differ only by a proline-rich 15-amino-acid sequence near the amino-terminus. In the increased expression with metastatic activity, the ratio of the isoform mRNAs remained constant, but the 929-amino-acid protein isoform showed increases up to about 3-fold in comparison to the 944-amino-acid isoform. The differential accumulation was explained by cellular labeling experiments that showed that the 944 isoform is degraded at a 5-fold greater rate than the 929 isoform and that this degradation required the ubiquitin-proteasome system.